This invention relates to a diagnostic method for the radioimmunoassay of antigens and their antibodies and a method for coating apparatus useful in the above determination. More particularly, this invention relates to a direct method for determining hepatitis associated antigens or their antibodies and is also directed toward a method for preparing diagnostic apparatus suitable for use in the same.
Although there have been methods for determining the presence of antigenically active macromolecules such as intact viruses, virus capsids, virus subunits, bacteria, membranes, cell walls, hormones, etc., there have been a lack of a simple, yet sensitive, test method and apparatus for determining the presence of these materials. Viral hepatitis, including so-called serum hepatitis, which is a relatively common disease, has not been heretofore easily detected by a sensitive test which is both specific and reproducible for quickly determining whether or not the sera from a patient or a donor contains hepatitis associated antigens or antibodies.
Furthermore, radioimmunoassay techniques have been developed in the past for various antigen-antibody materials; however, these radioimmunoassay techniques such as disclosed in articles by Keven Catt et al in the Journal of Biochemistry, 1966, Volume 100, pages 31c and 33c and in Science, Volume 158, page 1570, 167, are an indirect radioimmunoassay technique wherein the amount of antigen present in roughly inversely proportional to the amount of radiation emitted by the tracer material. These procedures required the use of correlation tables and other materials which generally rendered the results less than reproducible and exact. Wide, Karolinska Simposia, 1st Simposia, Sept. 23 to 25, 1969, University Hospital Upsila, Sweden, pages 207 to 214 discloses a radioimmunoassay wherein an allergen is absorbed on a plastic polymer and reagin is bound immunochemically to the solid phase allergen and then labeled anti-reagen is bound to the reagin.
Briefly, it has been discovered that the above-noted difficulties, i.e., lack of reproducibility and exactness, have been overcome by utilizing the method of the present invention. Briefly, in one application, the method of the present invention comprises contacting an unknown serum sample with an antibody coated implement, incubating the test implement and serum for a period of from 0.5 to 42 hours, aspirating and washing, contacting an I.sup.125 labled antibody with the serum and coated with the test apparatus and incubating for from 1 to 6 hours, aspirating and washing, and counting the tube for I.sup.125 gamma radiation.
It is therefore the principal object of the present invention to provide a novel method for the direct determination of antigens and their antibodies.
It is a further object of the present invention to provide a method for coating diagnostic apparatus for use in radioimmunoassay determinations.
It is a still further object of the present invention to provide a method for quickly and accurately determining the presence of hepatitis associated antigens or antibodies in sera.